Limb ischemic preconditioning protects human umbilical vein endothelial cells from oxidative stress induced by H2 O2 / 中国病理生理杂志

ABSTRACT

AIM: To investigate the effects of the sera from the rats after limb ischemic preconditioning (LIPC) on human umbilical vein endothelial cells (HUVECs) injured by hydrogen peroxide (H2O2).METHODS:The HUVECs were divided into 5 groupsthe cells in control group were cultured without any intervention;the cells in model group (M) were damaged by 1 mmol/L H2O2 for 2 h;the cells in early preconditioning serum (EPS) group, delayed pre-conditioning serum (DPS) group or sham limb ischemic preconditioning serum (SPS) group were treated with the corre-sponding serum at 5%for 12 h, respectively , and then treaed with H 2 O2 for 2 h.The viability of the HUVECs was ana-lyzed by flow cytometry .The lactate dehydrogenase ( LDH) in the culture media was detected .The cell adhesion molecules in the HUVECs were detected by real-time PCR.The mRNA and protein expression of heme oxygenase-1 (HO-1) was also determined .RESULTS:The viability of HUVECs incubated with 1 mmol/L H2 O2 for 2 h significantly decreased compared with the control cells , which was accompanied with the augmentations of LDH in the medium and the cell adhesion mole -cules in cells , such as vascular cell adhesion molecule-1 ( VCAM-1 ) and intercellular adhesion molecule-1 ( ICAM-1 ) . Preincubation with EPS and DPS derived from the rats subjected LIPC attenuated these injuries .Furthermore, pretreatment with EPS and DPS increased the expression of HO-1 at mRNA and protein levels .CONCLUSION:LIPC protects the HU-VECs from H2O2-induced injury.